Obtención de una columna monolítica de inmunoafinidad como contribución al desarrollo de un método de enriquecimiento de anticuerpos policlonales
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Insuasty Cepeda, Diego Sebastián
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Trabajo de grado - Maestría
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2018
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Los metabolitos tienen gran potencial en el diseño de biomarcadores ya que la presencia o ausencia de estos brinda información valiosa de un sistema biológico. En algunas patologías se ha encontrado que estos metabolitos se presentan en bajas concentraciones, en estadios tempranos o asintomáticos, por lo que es difícil detectarlos. Dentro de este contexto, un material monolítico orgánico de poli(GMA-co.EDMA) fue funcionalizado con un péptido por adición de Michael (tiol-maleimido, química click). El péptido, unido covalentemente al monolito, contiene la secuencia SPINNTKPHEAR derivada de la proteína L1 del Virus del Papiloma Humano (VPH). Se determinó, mediante ensayo de ELISA, que el soporte monolítico obtenido permite aislar selectivamente anticuerpos policlonales contra la secuencia SPINNTKPHEAR, ya que estos son retenidos en la superficie química del material mediante una interacción de inmunoafinidad. En este proyecto se desarrolló un nuevo método para aislar y preconcentrar metabolitos, específicamente anticuerpos, utilizando materiales monolíticos, lo que podría contribuir a la mejora de las estrategias de detección de enfermedades basadas en interacciones de inmunoafinidad.
Abstract: Metabolites have great potential in the biomarkers design since the presence or absence of these provides valuable information of a biological system. In some pathologies, it has been found that these metabolites are in low concentrations, in early or asymptomatic stages, so it is difficult to detect them. Here in, an organic monolithic material of poly(GMA-co-EDMA) was functionalized with a peptide by thiol-maleimide Michael addition click reaction. The covalently bound peptide, to the monolith, corresponds to the sequence SPINNTKPHEAR derived from the L1 protein of the Human Papilloma Virus (HPV). Using ELISA assays, it was determined that the obtained monolithic support allows to selectively isolated polyclonal antibodies against the sequence SPINNTKPHEAR, since these are retained in the chemical surface of the material by immunoaffinity interaction. In this project was developed a new strategy for isolating and preconcentrating metabolites, specifically antibodies, using monolithic materials, it could contribute to the improvement of diseases detection strategies based in inmunoaffinity interactions
Abstract: Metabolites have great potential in the biomarkers design since the presence or absence of these provides valuable information of a biological system. In some pathologies, it has been found that these metabolites are in low concentrations, in early or asymptomatic stages, so it is difficult to detect them. Here in, an organic monolithic material of poly(GMA-co-EDMA) was functionalized with a peptide by thiol-maleimide Michael addition click reaction. The covalently bound peptide, to the monolith, corresponds to the sequence SPINNTKPHEAR derived from the L1 protein of the Human Papilloma Virus (HPV). Using ELISA assays, it was determined that the obtained monolithic support allows to selectively isolated polyclonal antibodies against the sequence SPINNTKPHEAR, since these are retained in the chemical surface of the material by immunoaffinity interaction. In this project was developed a new strategy for isolating and preconcentrating metabolites, specifically antibodies, using monolithic materials, it could contribute to the improvement of diseases detection strategies based in inmunoaffinity interactions